PCR Detects the DNA Vaccines in mice Vaccinated via and Construction Containing Hsp70 and GP5 /M protein of PRRSV

Full Text (PDF, 348KB), PP.35-41

Views: 0 Downloads: 0

Author(s)

ZUO Lan 1 YAN Qi-gui 2,* CHEN Shi-jie 3 LEI Yan 1 WANG Xu 1

1. Animal Biotechnology Center, Sichuan Agricultural University, Ya’an, Sichuan, 625014,China

2. Key laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University,Ya’an,625014

3. Sichuan Entry-exit Inspection and Quarantine Bureau,Chengdu,610041,China

* Corresponding author.

DOI: https://doi.org/10.5815/ijieeb.2011.02.05

Received: 6 Nov. 2010 / Revised: 26 Dec. 2010 / Accepted: 10 Feb. 2011 / Published: 8 Mar. 2011

Index Terms

PRRSV, DNA vaccine, Hsp70, DNA immune, distribute

Abstract

Construction of Eukaryotic recombinant expression Vector pCI-ORF5-ORF6/ pCI-ORF5-ORF6B-Hsp70 Containing Hsp70 and PRRSV GP5/M (encoded by ORF5 and ORF6 genes), and to study its immune effect. After being identified by enzyme analysis and nucleotide sequencing test, the repression vector plasmid was transfected into COS-7 cells. The transient expression protein was detected by Western-blotting. The immunogenicities of this DNA vaccine constructs were firstly investigated in a mouse moder. IFN-γ, IL-4 of cytokine, and the spleen T-lymphocyte subgroup quantity (CD4+/CD8+) were detected, DNA vaccine distribution in mice by PCR .The result shows that the recombinant plasmid pCI-ORF5-ORF6-Hsp70 could induce higher response of cellular immune responses and specific immune responses in mouse, the DNA Vaccines in mice Vaccinated via as heart and liver ,lung and kidney, muscle and brain each time step after immunity. providing the clinical basic data and theoretical basis for success of the DNA vaccine development.

Cite This Paper

ZUO Lan, YAN Qi-gui, CHEN Shi-jie, LEI Yan, WANG Xu, "PCR Detects the DNA Vaccines in mice Vaccinated via and Construction Containing Hsp70 and GP5 /M protein of PRRSV", International Journal of Information Engineering and Electronic Business(IJIEEB), vol.3, no.2, pp.35-41, 2011. DOI:10.5815/ijieeb.2011.02.05

Reference

[1]WENSVOORT G, TERPSTRA C, POL J, et al. Mystery swine disease in The Netherlands: the isolation of Lelystad virus [J]. The Veterinary Quarterly, 1991, 13(3): 121.
[2]SHI-WEN T. Advances on Highly Pathogenic Porcine Reproductive and Respiratory Syndrome (PRRS)[J] [J]. Guizhou Agricultural Sciences, 2009, 6
[3]JIANG Y, FANG L, XIAO S, et al. A suicidal DNA vaccine co-expressing two major membrane-associated proteins of porcine reproductive and respiratory syndrome virus antigens induce protective responses [J]. Biotechnology letters, 2009, 31(4): 509-18.
[4]ZHOU Y, HAO X, TIAN Z, et al. Highly virulent porcine reproductive and respiratory syndrome virus emerged in China [J]. Transboundary and Emerging Diseases, 2008, 55(3‐4): 152-64.
[5]ZHOU Y, YU H, TIAN Z, et al. Monoclonal antibodies and conserved antigenic epitopes in the C terminus of GP5 protein of the North American type porcine reproductive and respiratory syndrome virus [J]. Veterinary microbiology, 2009, 138(1-2): 1-10.
[6]VOS M, HAGEMAN J, CARRA S, et al. Structural and Functional Diversities between Members of the Human HSPB, HSPH, HSPA, and DNAJ Chaperone Families [J]. Biochemistry, 2008, 47(27): 7001-11.
[7]SELVIN P, HA T. Single-molecule techniques: a laboratory manual [M]. Cold Spring Harbor Laboratory Pr, 2008.
[8]DURAN J, CLIMENT I, SARCASECA J. Baculovirus Expression of Porcine Reproductive and Respiratory Syndrome Virus Strain Olot/91: Involvement of ORF3 and ORF5 proteins in protection [J]. Virus genes, 1997, 14(19-29).
[9]PING-AN X, ZHAN-GUO D, BIN Z, et al. Pseudotyping of Murine Leukemia Virus Particles with Porcine Reproductive and Respiratory Syndrome Virus M Protein-mediated E Protein [J] [J]. Chinese Journal of Virology, 2008, 5.
[10]DAUGAARD M, ROHDE M, J TTEL M. The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions [J]. FEBS letters, 2007, 581(19): 3702-10.
[11]TSAN M, GAO B. Heat shock proteins and immune system [J]. Journal of leukocyte biology, 2009, 85(6): 905.
[12]ZHANG K, ZHAO T, HUANG X, et al. Preinduction of HSP70 promotes hypoxic tolerance and facilitates acclimatization to acute hypobaric hypoxia in mouse brain [J]. Cell Stress and Chaperones, 2009, 14(4): 407-15.
[13]KOKOLAKIS G, KRITSIDIMA M, TKACHENKO T, et al. Two hsp23 genes in the Mediterranean fruit fly, Ceratitis capitata: structural characterization, heat shock regulation and developmental expression [J]. Insect Molecular Biology, 2009, 18(2): 171-81.
[14]SINGH M, REDDY M, MATHUR N, et al. Induction of hsp70, hsp60, hsp83 and hsp26 and oxidative stress markers in benzene, toluene and xylene exposed Drosophila melanogaster: Role of ROS generation [J]. Toxicology and applied pharmacology, 2009, 235(2): 226-43.
[15]VAN DEN P, WOLLMANN G. COMPOSITIONS AND METHODS OF USE OF AN ONCOLYTIC VESICULAR STOMATITIS VIRUS [M]. US Patent App. 20,100/172,877.
[16]DAS S, LAXMINARAYANA S, CHANDRA N, et al. Heat shock protein 70 on Neuro2a cells is a putative receptor for Japanese encephalitis virus [J]. Virology, 2009, 385(1): 47-57.
[17]AN CHUN C, MING SHU W,NI GEN X,et al. Construction of ORF5 gene vaccine against PRRSV and detection of its safety and immunogenicity[J]. Chinese Journal of Veterinary Science and Technology,2004,35(1):27-35.
[18]WEN BO D,AN CHUN C, MING SHU W,et al .PCR Detects the PRRSV of ORF5 Gene Vaccine in BALB/c Mice Vaccinated via Gene Gun Bombing[C]. 2005.